US10403412B2 - Optical tweezers device - Google Patents

Optical tweezers device Download PDF

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Publication number
US10403412B2
US10403412B2 US15/770,781 US201615770781A US10403412B2 US 10403412 B2 US10403412 B2 US 10403412B2 US 201615770781 A US201615770781 A US 201615770781A US 10403412 B2 US10403412 B2 US 10403412B2
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particle
trapping force
distance
lens
laser beam
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US20180322976A1 (en
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Kota Nanri
Toshiyuki Saito
Kensuke Suzuki
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JTEKT Corp
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JTEKT Corp
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    • G—PHYSICS
    • G21—NUCLEAR PHYSICS; NUCLEAR ENGINEERING
    • G21K—TECHNIQUES FOR HANDLING PARTICLES OR IONISING RADIATION NOT OTHERWISE PROVIDED FOR; IRRADIATION DEVICES; GAMMA RAY OR X-RAY MICROSCOPES
    • G21K1/00—Arrangements for handling particles or ionising radiation, e.g. focusing or moderating
    • G21K1/006—Manipulation of neutral particles by using radiation pressure, e.g. optical levitation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B25—HAND TOOLS; PORTABLE POWER-DRIVEN TOOLS; MANIPULATORS
    • B25J—MANIPULATORS; CHAMBERS PROVIDED WITH MANIPULATION DEVICES
    • B25J7/00—Micromanipulators
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/32—Micromanipulators structurally combined with microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
    • G02B21/361—Optical details, e.g. image relay to the camera or image sensor
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • G02B21/08—Condensers
    • G02B21/086—Condensers for transillumination only

Definitions

  • An aspect of the present invention relates to an optical tweezers device.
  • the optical tweezers technology is known which is a technology for trapping, for example, a particle measuring about 1 ⁇ m and, furthermore, moving it (refer to Patent document 1, for example).
  • a laser beam is focused with a lens and a particle that is brought close to a focusing point is trapped by optical pressure acting on it.
  • the optical tweezers technology makes it possible to continue to trap a particle by providing a refractive index difference between the particle and what exists around it and directing the total force of optical pressure acting on the particle to the focusing point.
  • the particle have transmittivity (transmit the laser beam) and that the refractive index (n2) of the particle is larger than the refractive index (n1) of what exists around it (n2>n1).
  • Patent document 1 JP-A-2006-235319
  • a particle can be moved relative to liquid around it so as to follow a focusing point of a laser beam by causing a relative movement between the focusing point, that is, the focal point of a lens for focusing the laser beam, and a sample including the particle.
  • a particle trapping force originates from optical pressure of the laser beam, and it is known that the trapping force and the distance between the particle and the focal point of the lens (focusing point) have a linear relationship.
  • An object of an aspect of the invention is therefore to provide an optical tweezers device capable of trapping and moving a particle stably even in a case of moving the trapped particle in a region that is close to the surface of an object.
  • An optical tweezers device includes: a light source which emits a laser beam; a lens which focuses the laser beam emitted from the light source; a drive unit which moves, relative to each other, a particle trapped by focusing the laser beam with the lens and an object located in the vicinity of the particle; a detector which outputs a detection signal for determination of a distance between the trapped particle and a focal point of the lens; a trapping force calculation unit which determines trapping force data indicating a trapping force for the particle on the basis of the distance determined from the detection signal; a difference calculation unit which determines a difference between a trapping force theoretical value that is estimated according to a linear relationship between the distance between the trapped particle and the focal point of the lens and the trapping force for the particle and the trapping force indicated by the trapping force data; and an output control unit which controls a laser power of the light source on the basis of the difference of the trapping force.
  • the relationship between the trapping force for the particle being trapped and the distance between the particle and the focal point of the lens is linear in a region that is distant from the surface of the object, it is nonlinear and unpredictable in a region that is close to the surface of the object.
  • the trapping force for the particle can be compensated so that its relationship with the distance between the particle and the focal point of the lens comes close to a linear relationship. This makes it possible to trap and move the particle stably.
  • the difference calculation unit may convert the difference into a current value input to the light source that is correlated with a power of the laser beam emitted from the light source, and the output control unit may perform a feedback control for increasing or decreasing the current value.
  • This configuration facilitates acquisition of a desired trapping force by adjusting the power of the laser beam of the light source, and thereby makes it possible to trap and move a particle even more stably.
  • the aspect of the invention makes it possible to trap and move a particle stably and thereby prevent untrapping of the particle even in a case of moving the trapped particle in a region that is close to the surface of an object.
  • FIG. 1 is an explanatory diagram for description of the overall configuration of an optical tweezers device.
  • FIG. 2 is an image diagram for description of the function of a detector.
  • FIG. 3 is an explanatory diagram showing a particle to be trapped, lenses, etc.
  • FIGS. 4(A) and 4(B) are explanatory diagrams showing relationships between the particle trapping force and the particle-focal point distance.
  • FIG. 5 is a graph showing a relationship between the input current of a light source and the power of the laser beam of the light source.
  • FIG. 1 is an explanatory diagram for description of the overall configuration of an optical tweezers device 1 .
  • the optical tweezers device 1 includes a light source 10 of a laser beam, light guiding means ( 21 - 27 ), a first lens 28 , an illumination light source 30 , a second lens 31 , a mirror (third mirror) 33 , a detector 40 , a device base 45 , a stage 46 , a drive means 48 , an imaging means 50 , and a control means 60 .
  • the optical tweezers device 1 is configured in such a manner that the stage 46 can be moved by the drive means 48 with respect to the device base 45 which is fixed to a working floor. And other devices, that is, the light source 10 , the lenses 28 and 31 , the detector 40 , the imaging means 50 , etc., are fixed to the device base 45 and are not moved with respect to the device base 45 .
  • the light source 10 of the laser beam which is a laser device for emitting the laser beam L, emits the laser beam L having a first wavelength according to a control signal received from the control means 60 .
  • a particle being held by a holding member (e.g., prepared slide) 47 mounted on the stage 46 is trapped by an optical tweezers technique using the laser beam L (optical trap).
  • the light guiding means ( 21 - 27 ) serve to guide the laser beam L emitted from the light source 10 to the first lens 28 .
  • the light guiding means ( 21 - 27 ) will be described below in order.
  • a first reflection mirror 21 reflects the laser beam L coming from the light source 10 so that it is incident on a first aperture 22 .
  • the first aperture 22 narrows the diameter of the incident laser beam L and outputs resulting light toward a first collimating lens 23 .
  • the first collimating lens 23 enlarges the diameter of the laser beam L and outputs resulting light toward a second collimating lens 24 .
  • the second collimating lens 24 converts the diameter-enlarged laser beam L into parallel light and outputs it to a second aperture 25 .
  • the second aperture 25 narrows the diameter of the parallel laser beam L and outputs resulting light toward a first mirror 26 .
  • the first mirror 26 reflects the incident laser beam L toward a second mirror 27 .
  • the second mirror 27 reflects the incident laser beam L toward the first lens 28 .
  • the first lens 28 focuses the laser beam L coming from the second mirror 27 at a focal point that is set in the holding member 47 .
  • a particle that is brought close to the focusing point (the focal point of the lens 28 ) can be trapped by the laser beam L focused by the lens 28 .
  • the focused laser beam L is incident on the second lens 31 after passing through the particle.
  • the laser beam L incident on the second lens 31 after passing through the particle is output toward the third mirror 33 , is reflected by the third mirror 33 , and is incident on the detector 40 .
  • the mirrors 33 and 26 transmit illumination light S coming from the illumination light source 30 .
  • the illumination light source 30 which is, for example, an LED illumination device, emits illumination light S having a second wavelength according to a control signal received from the control means 60 .
  • the illumination light S serves as illumination light for the imaging means 50 for observing a state of a particle being held by the holding member 47 .
  • the illumination light S passes through the third mirror 33 , is focused by the second lens 31 , thereafter passes through the first lens 28 , is reflected by the second mirror 27 , passes through the first mirror 26 , and reaches the imaging means 50 .
  • the holding member 47 for holding a particle is mounted on the stage 46 .
  • a fluid W and a particle C to be trapped that is contained in the fluid W are held by the holding member 47 (see FIG. 3 ).
  • An object B not to be trapped by the laser beam L is also held by the holding member 47 .
  • the object B and the trapped particle C move relative to each other.
  • the fluid W may contain particles (not shown) not to be trapped.
  • the fluid W is a liquid.
  • the refractive index (n1) of the fluid W is smaller than the refractive index (n2) of the particle C (n1 ⁇ n2).
  • the stage 46 is supported so as to be movable in the front-rear direction, the left-right direction, and the top-bottom direction, and the drive means 48 moves the stage 46 in the front-rear direction, the left-right direction, and the top-bottom direction.
  • the X-axis direction, the Y-axis direction, and the Z-axis direction are defined as the front-rear direction, the left-right direction, and the top-bottom direction, respectively.
  • the drive means 48 moves the stage 46 in at least one of the X-axis direction, the Y-axis direction, and the Z-axis direction according to a control signal received from the control means 60 and thereby moves the holding member 47 in the same direction.
  • the drive means 48 is composed of actuators using a piezoelectric element, for example. In the embodiment, a description will be made of a case that the stage 46 is moved in the X-axis direction in the XY plane.
  • the focusing point (focal point of the lens 28 ) is not moved by the drive means 48 .
  • a particle C that has come close to the focusing point (focal point) and is trapped is not moved by the drive means 48 .
  • the stage 46 is moved together with the holding member 47 by the drive means 48 .
  • fluid W around the particle C that has come close to the focusing point and is trapped see FIG. 3
  • the object B and the particles contained in the fluid W and not trapped
  • the detector 40 is a position detector for detecting an incident position of the laser beam L with respect to a reference position; in the embodiment, the detector 40 is a quadrant position detector.
  • FIG. 2 is an image diagram for description of the function of the detector 40 .
  • the detector 40 has photodetection units A 1 , A 2 , A 3 , and A 4 obtained by dividing a flat surface into plural (four) portions.
  • FIG. 3 is an explanatory diagram showing the particle C to be trapped, the lens 28 , etc. Coordinates in the XY plane including the focal point Q of the lens 28 (see FIG. 3 ) are correlated with XY-plane coordinates of the photodetection units A 1 , A 2 , A 3 , and A 4 (see FIG.
  • Each of the photodetection units A 1 , A 2 , A 3 , and A 4 outputs a detection signal (voltage signal) corresponding to a receiving position J of the laser beam L.
  • the holding member 47 including the particle C being trapped is moved in the X-axis direction with respect to the lens 28 (see FIG. 3 ), the particle C is moved in the X-axis direction (in the direction opposite to the movement direction of the holding member 47 ) so as to follow the focal point Q of the lens 28 .
  • the laser beam L that reaches the photodetection units A 1 , A 2 , A 3 , and A 4 after being emitted from the light source 10 and passing through the particle C being trapped is detected at a position that is deviated in the X-axis direction from the reference position N (see FIG. 2 ) corresponding to the position of the focal point Q of the lens 28 by a distance d 0 .
  • the detector 40 is a quadrant position detector, the distance d 0 is output in the form of a voltage (voltage signal) (V).
  • the distance d 0 (V) correlates with the distance d (m) (see FIG. 3 ) between the center position of the particle C being trapped by the laser beam L and the focal point Q of the lens 28 ; distances d 0 (V) are output one after another. That is, detection signals (voltage signals) (V) to be used for determination of distances d between the particle C being trapped and the focal point Q of the lens 28 are output from the detector 40 one after another.
  • These detection signals are input to the control means (computer; described later) 60 and processed by a distance calculation unit 61 (see FIG. 1 ) provided in the control means 60 , whereby distances d between the particle C being trapped and the focal point Q of the lens 28 are determined one after another.
  • These distances d are values in the XY plane. The distance d also varies because the distance d 0 varies depending on the movement velocity of the stage 46 .
  • the imaging means 50 shown in FIG. 1 which is a CCD camera or a CMOS camera, for example, images a region including the focusing point and its neighborhood.
  • the imaging means 50 outputs image data produced by imaging to the control means 60 .
  • the control means 60 which is, for example, a computer that is equipped with a processor and a memory, outputs control signals as described above and takes in image data supplied from the imaging means 50 .
  • the control means 60 is equipped with the distance calculation unit 61 , a trapping force calculation unit 62 , a difference calculation unit 63 , and an output control unit 64 as function units that are implemented by the processer's running computer programs that are stored in the memory of the computer.
  • the distance calculation unit 61 determines, through calculation, a distance d (see FIG. 3 ) between the particle C being trapped and the focal point Q of the lens 28 on the basis of a detection signal (voltage signal) received from the detector 40 .
  • a detection signal voltage signal
  • the distance d between the particle C being trapped and the focal point Q of the lens 28 will be referred to as a “particle-focal point distance d.”
  • the calculation of a distance d by the detector 40 and the distance calculation unit 61 can be performed by means that are employed conventionally in the optical tweezers technology. Example processing for calculating a distance d will be described below. As described above, distances d 0 (V) are detected by the detector 40 one after another.
  • R in this equation is a value that was determined in advance by another piece of processing.
  • the value of R is determined in the following manner. For example, a particle that is fixed to the holding member 47 is caused to traverse (pass through) the laser beam (focal point) at a constant velocity. At this time, the particle is not trapped by the laser beam.
  • This constant-velocity manipulation is performed using voltages V that are supplied to the piezoelectric element of the drive means 48 .
  • the coefficient h in this equation is known from the characteristic of the detector 40 .
  • the trapping force calculation unit 62 determines trapping force data indicating trapping forces for the particle C on the basis of the particle-focal point distances d that have been determined by the distance calculation unit 61 from the detection signals of the detector 40 . Processing for determining trapping force data will be described later.
  • FIGS. 4(A) and 4(B) are explanatory diagrams showing relationships between the trapping force Tx for the particle C and the particle-focal point distance d.
  • the vertical axis represents the trapping force Tx for the particle C
  • the horizontal axis represents the particle-focal point distance d.
  • FIG. 4(A) shows a case that the particle C being trapped moves in a region that is distant from the surface of the object B
  • FIG. 4(B) shows a case that the particle C being trapped moves in a region that is close to the surface of the object B.
  • the distance d and the trapping force Tx have a linear relationship (proportional relationship).
  • the distance d and the trapping force Tx have a linear relationship in a range in which the distance d is short but have a nonlinear (irregular) relationship in a range in which the distance d is greater than a certain value.
  • k is a constant (a spring constant of optical trapping)
  • f is the external force.
  • the difference calculation unit 63 determines a difference (Txi ⁇ Txa) between a trapping force theoretical value Txi that is estimated according to a linear relationship that holds between the particle-focal point distance d and the trapping force Tx for the particle C (obtained in a region that is distant from the surface of the object B) and a trapping force Txa indicated by the trapping force data determined by the trapping force calculation unit 62 .
  • the parameter ⁇ in this equation is influenced by, among other things, static electricity and formation of a standing wave due to reflection of the laser beam, and is a variable.
  • a feedback control is performed in which the laser power control by the output control unit 64 is repeated as time elapses on the basis of ⁇ T ( ⁇ ).
  • the straight lines shown in FIGS. 4(A) and 4(B) have the same constant k.
  • the relationships (data) shown in FIGS. 4(A) and 4(B) can be determined on the basis of experimental values or calculation values.
  • the light source 10 employed in the embodiment uses a semiconductor laser and, as shown in FIG. 5 , the input current I of the semiconductor laser and the power P of the laser beam L emitted from the light source 10 have a proportional relationship. Furthermore, the power P of the laser beam L and the particle trapping force T have a proportional relationship as indicated by the following Equation (1).
  • Equation (1) a is a coefficient that is attributed to the refractive index and the transmittance of a particle, the refractive index of liquid (solvent) around it, and the beam waist of laser beam L.
  • the current I and the trapping force T also have a proportional relationship.
  • the relationship between the particle-focal point distance d and the trapping force T can be made close to a proportional relationship. This makes it possible to prevent untrapping of the particle C by securing trapping forces T that are suitable for particle-focal point distances d.
  • the optical tweezers device 1 may have another configuration. That is, the optical tweezers device 1 is equipped with the light source 10 for emitting the laser beam L, the lens 28 for focusing the laser beam L emitted from the light source 10 , the drive means 48 for moving, relative to each other, the lens 28 and the stage 46 which is mounted with the holding member 47 including the particle C to be trapped, the detector 40 for outputting a detection signal for determination of a distance d between the particle C being trapped and the focal point Q of the lens 28 , and the control means 60 (computer) for performing various kinds of processing.
  • the control means 60 computer
  • the holding member 47 is provided with, in addition to the liquid W, the object B whose relationship with the particle C to be trapped is to be examined (see FIG. 3 ).
  • the drive means 48 can move, relative to each other, the particle C trapped by focusing the laser beam L by the lens 28 and the object B located in the vicinity of the particle C.
  • the particle C is moved in a region that is close to the surface of the object B parallel with its surface.
  • the relationship between the trapping force T for the particle C being trapped and the particle-focal point distance d is linear in a region that is distant from the surface of the object B, it is nonlinear and unpredictable in a region that is close to the surface of the object B.
  • the trapping force T for the particle C can be compensated so that its relationship with the particle-focal point distance d comes close to a linear relationship. This makes it possible to trap and move the particle C stably. That is, even in the case of trapping and moving the particle C in a region that is close to the surface of the object B, untrapping of the particle C can be prevented.
  • This configuration facilitates acquisition of a desired trapping force T by adjusting the power of the laser beam L of the light source 10 , and thereby makes it possible to trap and move a particle C even more stably.
  • optical tweezers device according to the invention is not limited to the one in the illustrated embodiment and may be one in any other embodiment within the scope of the invention.
  • the optical tweezers device 1 By trapping a particle and moving it relative to an object using the optical tweezers device 1 , it becomes possible to measure a surface shape of the object, measure a surface force involving the object, or perform microprocessing on the object. Furthermore, the viscosity or the like of liquid W around an object B can be detected by determining a trapping force for a particle.

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CN109920575B (zh) * 2019-03-20 2020-08-25 中国人民解放军国防科技大学 一种基于二维光阱的自冷却激光光镊装置和方法

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DE112016004958T5 (de) 2018-07-12
JP6606975B2 (ja) 2019-11-20
US20180322976A1 (en) 2018-11-08
JP2017083644A (ja) 2017-05-18
WO2017073470A1 (ja) 2017-05-04
CN108349075B (zh) 2021-04-30

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